Flow-cytometric light scattering measurement of red blood cell volume and hemoglobin concentration

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Abstract
A light scattering technique for simultaneously determining the volume (V) and hemoglobin concentration (HC) of individual sphered red blood cells (RBCs) is described. Light scattered into two angular intervals yields measurements S1 and S2, respectively. Since a sphered RBC is essentially a homogeneous dielectric sphere having a complex refractive index that is linear in HC, with a proper choice of detector acceptance angles, tables relating V and HC to S1 and S2 can be computed via Mie theory. Absolute calibration is possible using droplets of water-immisible oils of accurately known refractive index. Results of experimental tests of the method are compared with those obtained from hematological reference measurements.